Drift-Diffusion Analysis of Neutrophil Migration during Inflammation Resolution in a Zebrafish Model

1Department of Automatic Control and Systems Engineering, University of Sheffield, Sheffield S1 3JD, UK2MRC Centre for Developmental and Biomedical Genetics, University of Sheffield, Firth Court, Western Bank, Sheffield S10 2TN, UK3University of Sussex School of Engineering and Design Biomedical Engineering Research Group, Brighton BN1 9QT, UK4Department of Infection and Immunity, University of Sheffield, Sheffield S10 2JF, UK

Supplementary Material

Supplementary Mathematical Methodology: The model equations are derived via solution of the drift‐diffusion equation subject to appropriate boundary conditions. Also described is the weighted least squares analysis which is adapted to identify the model parameters and associated uncertainties for the incoming (green labelled) cells and outgoing (red labelled) cells respectively.

Supplemental Movie: 3 dpf embryos from transgenic zebrafish expressing Kaede in neutrophils were subjected to tailfin transection under anaesthesia using a sterile scalpel. The embryos were
recovered for 4 hours. At four hours after injury the embryo was mounted in 0.5% low melting point agarose for imaging on a Laser Confocal System (Perkin Elmer Inc). The PhotoKinesis device was then used to photoconvert all neutrophils present within the tip of the tailfin. Photoconversion was carried out according to the methods described (120 cycles of 40% 405 nm laser energy), timelapse videomicroscopy was performed using a TE200U fluorescent inverted microscope (Nikon).

The timelapse sequence represents the full dataset shown in Figure 1 and 2. Composite images of DIC overlaid with the red and green fluorescence channels.